*edit 12/29/21.. link deleted, was for an unrelated book*

This procedure was replicated by sonication @ 80W, of a dry gram of p.cubensis aborts in ~50 mL hplc-grade methanol for 10 mins at 25 C , followed by magnetic stirring for 24 hr. The solution was filtered through 0.44 um syringe filters into 1.5 mL autosampler vials. HPLC-UV analysis was conducted isocratically, the mobile phase consisted of 10:90:1 MeOH:H2O:AcOH, at 333 uL/min. @ 25C. Excitation wavelength was 283 nm. The column used was a Phenomenex Kinetex C18 100x2.1mm, 2.6um 100Å.

The implications are clear. This is a potency/purity assay that would be very useful for biotransformation studies.

The bottom chromatogram is an ongoing investigation, as six fractions were collected for further analysis. Separation was done with a ramp gradient from 100% of the aforementioned mobile phase, to 100% of 0.1% Formic acid in Acetonitrile, over the course of 10 mins, holding at the latter mobile phase for the duration.

The same mobile phase was used, at 0.3 mL/min. The samples, 0.5g of controls and 0.5g experimental (10mM tryptamine hcl) aborts, were homogenized in 10mL hplc-grade methanol, and centrifuged @ 5000rpm for 5 min. Aliquots of 100uL from each were diluted with 900uL MeOH; 10uL injections were analyzed, 286nm excitation.

the first three are controls, the second three are experimental; notated in area counts. Taking the mean of the counts, the experimental group resulted in 6% increase in psilocybin.

today 1.5mL aliquots from the stock sol'n were taken, and the result was a 9.125% difference between the respective area counts.

there appears to be a direct correlation to the addition of dopant, and the augmentation of area counts. Further experiments will be conducted with 15mM tryptamine hcl.

Benzyme--

You are in waters way beyond my depth, but I may have bumped into a paper that would be germane to your work. "Determination of Psilocybin and Psilocin in Hallucinogenic Mushrooms by HPLC with Diode Array and MS Detection"(2008 )

Here is a link to the article, and I'll try to attach it below.

https://www.thevespiary.org/rhod...and-MS-Detection1b3b.pdf

I've been trying to untangle the myths around the lemon tek, and am amazed at how little is known about the quantities of psilocin and psilocybin in the mushroom or dephosphorylation of psilocybin to psilocin.

Hope this is at least a bit helpful. I greatly appreciate your work.

thank you, sir.

after doing some reading, I discovered that the small peak that co-elutes with the large peak is baeocystin.

Thank-you for your work Benzyme.

You're most welcome. It's what I do when I'm not 'at work'.

What about psiclocin content? Gartz found a huge boost to psiclocin content when he doped with 25mM of tryptamine HCl.

I love this work by the way. Thank you for doing it benzyme.

I didn't do the math, but the second peak is psilocin, and there is an increase in counts. It's only 10mM supplementation.

Right, it's just interesting that at 25mM a very large psiclocin increase was seen back in 1989.

Back to roost, with a more sensitive detector (fluorescence detector) and fancier software.
Attached is a typical HPLC chromatogram of dried psilocybe sample in methanol, fluorescence detection, and a 3D contour plot, Emission scan (wavelength vs. time vs. intensity (counts)).

What stands out to me is the second smaller peak has a larger emission lambda max (434.75 nm), which is where the cursor line on the right is positioned . The primary peak's lambda max is notated. This tells me there is a greater degree of pi-bond conjugation in that secondary peak.

I just posted a paper in the science section, which elucidated that beta-carbolines are present in psilocybe spp. That's not likely what this peak is (possibly baeocystin).

Speaking out of ignorance: Do you think pi bonding helps psilocin molecules come closer and begin oxidation at higher (but moderate) pH ~ pI? Asking, because if yes, adding a salt with K+ may help disrupt the process in water and help the poor yields in ongoing experimental kitchen test extractions. Of course, one can always just try, but wondering what you think.

that's a reasonable postulation; I don't know for certain, I just know that hydrogen on the hydroxy gets plucked off around a pH of 9, leaving O-, which is particularly unstable.

a colleague is going to investigate liposomal formulations; it would make sense to incorporate dilute ascorbic acid in the formulation.

Psilo research will be my particular focus, and I may collab with said colleague, who also has instrumentation.

Looking to hear more about this